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Structure of a cation-bound multidrug and toxic compound extrusion transporter

Abstract

Transporter proteins from the MATE (multidrug and toxic compound extrusion)1 family are vital in metabolite transport in plants2,3, directly affecting crop yields worldwide4. MATE transporters also mediate multiple-drug resistance (MDR) in bacteria and mammals5, modulating the efficacy of many pharmaceutical drugs used in the treatment of a variety of diseases6,7,8,9. MATE transporters couple substrate transport to electrochemical gradients and are the only remaining class of MDR transporters whose structure has not been determined10. Here we report the X-ray structure of the MATE transporter NorM from Vibrio cholerae determined to 3.65 Å, revealing an outward-facing conformation with two portals open to the outer leaflet of the membrane and a unique topology of the predicted 12 transmembrane helices distinct from any other known MDR transporter. We also report a cation-binding site in close proximity to residues previously deemed critical for transport11. This conformation probably represents a stage of the transport cycle with high affinity for monovalent cations and low affinity for substrates.

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Figure 1: NorM-VC model.
Figure 2: Validation of the NorM-VC model, showing the positions of the 16 mercury-labelled cysteine mutants.
Figure 3: Electrostatic potential surface representation of NorM-VC.
Figure 4: The cation-binding site of NorM-VC and mechanism of transport.

Accession codes

Primary accessions

Protein Data Bank

Data deposits

Coordinates and structure factors for the structures reported here are deposited in the Protein Data Bank with accession codes 3MKT and 3MKU.

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Acknowledgements

We thank Y. Yin for his contribution to initial clones of NorM. We also thank the Canadian Light Source (CLS), the Stanford Synchrotron Radiation Laboratory (SSRL), the Advanced Light Source (ALS) and the Advanced Photon Source (APS). This work was supported by grants from the National Institutes of Health (GM70480 to G.C., and GM73197 to Q.Z.), the Beckman Foundation and the Skaggs Chemical Biology Foundation.

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X.H., P.S., A.K. and G.C. designed the experiments and wrote the manuscript. X.H., P.S., A.K. and R.E. performed experiments. W.H. and Q.Z. did chemical synthesis.

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Correspondence to Geoffrey Chang.

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The authors declare no competing financial interests.

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He, X., Szewczyk, P., Karyakin, A. et al. Structure of a cation-bound multidrug and toxic compound extrusion transporter. Nature 467, 991–994 (2010). https://doi.org/10.1038/nature09408

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